Establishment And Charactetizaton Of Fin Cell Lines From Diploid, Triploid, And Tetraploid Oriental Weatherfish (Misgurnus Anguillicaudatus)

Oriental weatherfish(Misgurnus anguillicaudatus)is one of the most popular culturedspecies in China, Korean peninsula and Japanese archipelago. Primary culture of fin fromdiploid,triploid and tetraploid oriental weatherfish was performed with tissue explant method.Until now, cells of three tissues have been subcultured to passage84for diploid,75for triploid,and68for tetraploid, respectively. The special and combined aseptic processing method wasimmersing the fin tissue in10%povidone-iodine for15min, and then in penicillin andstreptomycin mixture (500IU/mL penicillin and500μg/mL streptomycin) for30min. Themedium of primary culture and early subculture was DMEM/F12, supplementing with20%fetal bovine serum(FBS),10ng/mL basic fibroblast growth factor（bFGF）,20ng/mL insulin-likegrowth factor-I（IGF-I）and10μg/mL chondroitin sulfate(pH7.2). The medium after passage30was only20%FBS-DMEM/F12. The condition of cell culture was25℃with5%CO2continuously. The feature chromosome number collected from100metaphase plates were2n=50,3n=75,4n=100in diploid,triploid and tetraploid fin cells, respectively.At the same time,the resultof SSR prove that the three cell lines are from oriental weatherfish.The diploid,triploid andtetraploid fin cells are named as DIMF,TRMF and TEMF, respectively.The doubling time were48.43h、36.01h and41.45h in diploid,triploid and tetraploid fincells at the50th passage. The cell livability of these three kinds of cell were（80.88±1.38）%，（84.48±1.13）%and（81.57±1.28）%when recovered after being stored in liquid nitrogen for60d at the40th passage. The feature chromosome number collected from100metaphase plateswere2n=50,3n=75in diploid,triploid fin cells at passage60, respectively,but the featurechromosome number was4n=96at the passage60. Establishment of fin cell lines from differentploidy oriental weatherfish in this study enriched the kinds of the fish cell lines, and would laythe foundation for discovering the mechanism of growth and genetics in polyploidy fish.